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目的:从前额叶皮质、海马及下丘脑中5-羟色胺(5-hydroxytryptamine,5-HT)和5-羟色胺受体3(5-hydroxytryptamine receptor3,5-HTR3),5-羟色胺受体4(5-HTR4)角度探讨健脾化湿颗粒改善腹泻型肠易激综合征(diarrhea-predominant irritable bowel syndrome,D-IBS)模型大鼠结肠运动和内脏敏感性的作用机制.方法:采用番泻叶灌胃结合束缚应激法建立D-IBS大鼠模型,应用健脾化湿颗粒进行干预,采用酶联免疫法(ELISA)检测大鼠海马中5-HT含量,采用免疫组织化学法检测前额叶皮质、海马及下丘脑中5-HT、5-HTR3、5-HTR4阳性表达,采用逆转录-聚合酶链反应法检测海马中5-H T R3 m R N A和5-H T R4m RNA的表达水平.结果:与正常组相比,模型组海马中5-HT含量(327.30±22.35 vs 265.33±13.60),前额叶皮质、海马、下丘脑中5-HT阳性表达(0.16±0.02 vs 0.08±0.01,0.19±0.02 vs 0.09±0.01,0.17±0.02 vs 0.08±0.01)明显升高(P<0.01);前额叶皮质、海马、下丘脑中5-HTR3阳性表达(0.29±0.02 vs 0.10±0.01,0.23±0.02 vs 0.09±0.01,0.22±0.02 vs 0.09±0.02)及5-HTR4阳性表达(0.25±0.02 vs0.11±0.01,0.28±0.02 vs 0.10±0.02,0.27±0.02 vs 0.11±0.02)明显升高(P<0.01);海马中5-H T R3 m R N A和5-H T R4 m R N A的表达(0.54±0.01 vs 0.17±0.05,0.73±0.08 vs 0.10±0.02)显著升高(P<0.01).与模型组相比,阳性对照组、中、高剂量组海马中5-H T含量(298.92±12.16、286.29±24.43、279.86±20.05 vs 327.30±22.35)显著下降(P<0.05,P<0.01),中、高剂量组前额叶皮质中5-HT表达(0.12±0.01、0.11±0.01 vs 0.16±0.02)显著下降(P<0.01),阳性对照组、中、高剂量组海马、下丘脑中5-H T表达显著下降(P<0.05,P<0.01);各治疗组前额叶皮质、海马、下丘脑中5-H T R3表达及5-H T R4表达下降显著(P<0.05,P<0.01);各治疗组海马中5-H T R3 m R N A表达及5-H T R4 m R N A表达显著降低(P<0.05,P<0.01).结论:健脾化湿颗粒可能通过下调脑中5-HT、5-HTR3、5-HTR4表达来改善D-IBS模型大鼠结肠运动和内脏敏感性.
Objective: To investigate the effects of 5-hydroxytryptamine (5-HT) and serotonin receptor 5 (5-HTR3) on the cortex, -HTR4) in order to explore the mechanism of action of “Jianpi Huashi Granule” on colonic motility and visceral sensitivity in diarrhea-predominant irritable bowel syndrome (D-IBS) model.Methods: Stomach combined with restraint stress method to establish D-IBS rat model, with the spleen and dampness granules for intervention, using enzyme-linked immunosorbent assay (ELISA) rat hippocampus 5-HT content, using immunohistochemistry to detect prefrontal cortex , 5-HT and 5-HTR4 in the hippocampus and hypothalamus, and the expression of 5-HT3R m RNA and 5-HT R4m RNA in the hippocampus were detected by reverse transcription-polymerase chain reaction.Results: Compared with the normal group, the 5-HT content (327.30 ± 22.35 vs 265.33 ± 13.60), the positive expression of 5-HT in the prefrontal cortex, the hippocampus and the hypothalamus of the model group (0.16 ± 0.02 vs 0.08 ± 0.01, 0.19 ± 0.02 vs 0.09 ± 0.01,0.17 ± 0.02 vs 0.08 ± 0.01, respectively); prefrontal cortex, hippocampus, hypothalamus 5-HTR3 positive expression (0.29 ± 0.02 vs 0.10 ± 0.01,0.23 ± 0.02 vs 0.09 ± 0.01,0.22 ± 0.02 vs 0.09 ± 0.02) and 5-HTR4 positive expression (0.25 ± 0.02 vs0.11 ± 0.01,0.28 ± 0.02 vs 0.10 ± 0.02,0.27 ± 0.02 vs 0.11 ± 0.02) (P <0.01). The expressions of 5-HT3 mRNA and 5-HT R4 mRNA in the hippocampus were significantly higher than those in the control group (0.54 ± 0.01 vs 0.17 ± 0.05, 0.73 ± 0.08 vs 0.10 ± 0.02) (P <0.01). Compared with the model group, the content of 5-HT in the hippocampus of the positive control group, middle and high dose group (298.92 ± 12.16,286.29 ± 24.43,279.86 ± 20.05 vs 327.30 ± 22.35) (P <0.05, P <0.01). The expression of 5-HT in the prefrontal cortex was significantly lower in the middle and high dose groups (0.12 ± 0.01,0.11 ± 0.01 vs 0.16 ± 0.02) (P <0.01) The 5-HT expression in the hippocampus and hypothalamus of the rats in the middle, high and high dose groups was significantly decreased (P <0.05, P <0.01). The expression of 5-HT3 in the cortex, hippocampus and hypothalamus in the prefrontal cortex and the 5-HT R4 (P <0.05, P <0.01). The expression of 5-HT3 mRNA and the expression of 5-HT4 mRNA in the hippocampus of each treatment group were significantly decreased (P <0.05, P <0.01) Wet particles may improve the expression of 5-HT, 5-HTR3, 5-HTR4 in the brain by down-regulating the expression of D- Bowel Movement and Visceral Sensitivity in IBS Model Rats.